Image enhancement method and widefield fluorescence imaging system

Abstract

A widefield fluorescence imaging system, comprising: arranging fluorescent microspheres on an image plane of widefield fluorescence microscopy, and collecting, by the widefield fluorescence microscopy, the microsphere images of the fluorescent microspheres; processing, by the computer equipment, the microsphere image, to obtain the half-peak full width value of the fluorescent microspheres; constructing a point diffusion model based on the half-peak full width value in computer equipment; collecting a sample image by the widefield fluorescence microscope, and then processing, by the computer device, the sample image based on a point diffusion model to obtain an enhanced image. The beneficial effect is that by detecting the half-peak full width value of the fluorescent microspheres and using it in the construction process of the point diffusion model, the processing process is simplified and the processing efficiency is improved while achieving better imaging results.

What is claimed is: 
     
         1 . An image enhancement method for widefield fluorescence imaging system comprising a widefield fluorescence microscope and a computer equipment connected to the widefield fluorescence microscope, the method comprising:
 arranging fluorescent microspheres on an image plane of the widefield fluorescence microscope, and collecting, by the widefield fluorescence microscope, the microsphere image of the fluorescent microspheres;   processing, by the computer equipment, the microsphere image, to obtain the half-peak full width value of the fluorescent microspheres;   constructing a point diffusion model based on the half-peak full width value in the computer equipment;   collecting a sample image by the widefield fluorescence microscope, and then processing, by the computer equipment, the sample image based on the point diffusion model to obtain an enhanced image.   
     
     
         2 . The image enhancement method according to  claim 1 , wherein before arranging the fluorescent microspheres, the widefield fluorescence microscope collects a background image on the image plane;
 after collecting, by the widefield fluorescence microscope, the microsphere image, the computer equipment outputs the microsphere image after noise removal based on the background image.   
     
     
         3 . The image enhancement method according to  claim 1 , wherein in the process of collecting microsphere image by the widefield fluorescence microscope, the widefield fluorescence microscope collects a plurality of fluorescence images on the fluorescent microspheres in turn;
 the computer equipment receives each of the fluorescence images in turn;   the computer equipment averages all of the fluorescence images and outputs them to function as the microsphere image.   
     
     
         4 . The image enhancement method according to  claim 1 , wherein the process of processing the microsphere image by the computer equipment further comprises:
 performing morphological processing, by the computer equipment, on the microsphere image to obtain a preprocessed image;   screening, by the computer equipment, the fluorescent microspheres from the preprocessed image, to obtain target microspheres;   generating, by the computer equipment, the half-peak full width value according to the image data of the target microspheres.   
     
     
         5 . The image enhancement method according to  claim 4 , wherein the process of screening, by the computer equipment, the fluorescent microspheres further comprises:
 performing, by the computer equipment, a detection of connectivity domain on the preprocessed image to obtain a plurality of connectivity domains, and removing, according to the connectivity domain, the adjacent fluorescent microspheres;   generating, by the computer equipment, a roundness of each of the fluorescent microspheres after screening;   comparing, by the computer equipment, the roundnesses with a preset roundness threshold to remove the fluorescent microspheres whose roundness is less than the roundness threshold;   outputting, by the computer equipment, the screened fluorescent microspheres to function as the target microspheres.   
     
     
         6 . The image enhancement method according to  claim 3 , wherein the process of generating the half-peak full width value by the computer equipment according to the image data comprises:
 performing, by the computer equipment, a nonlinear least squares fitting to the image data of each of the target microspheres to obtain the half-peak full width value of the fluorescent microspheres;   averaging, by the computer equipment, all of the half-peak full width value of the fluorescent microspheres to obtain the half-peak full width value.   
     
     
         7 . The image enhancement method according to  claim 1 , wherein the process of processing the sample image by the computer equipment comprises:
 processing, by the computer equipment, the sample image based on the point diffusion model to obtain an intermediate image of this iteration;   generating, by the computer equipment, an evaluation result of this iteration according to the intermediate image, the point spread function, and the sample image;   determining, by the computer equipment, whether the intermediate image meets a preconfigured iteration condition;   if yes, outputting, by the computer equipment, the intermediate image to function as the enhanced image;   If not, processing, by the computer equipment, the intermediate image based on the point diffusion model to obtain an intermediate image of next iteration.   
     
     
         8 . A widefield fluorescence imaging system applying the image enhancement method according to  claim 1 , comprising:
 the widefield fluorescence microscope for collecting the sample image;   the computer equipment connected to the widefield fluorescence microscope, the computer equipment receives the sample image and processes the sample image based on the image enhancement method to obtain the enhanced image.   
     
     
         9 . The widefield fluorescence imaging system according to  claim 8 , wherein the computer device comprises:
 a model building module, the model building module constructs the point diffusion model according to the microsphere image collected by the widefield fluorescence microscope;   the image enhancement module, the image enhancement module is connected to the model building module, and the image enhancement module processes the sample image according to the point diffusion model to obtain the enhanced image.   
     
     
         10 . The widefield fluorescence imaging system according to  claim 9 , wherein the image enhancement module is provided with an iterative submodule, the iterative submodule iterates the sample image according to the point diffusion model, to obtain the enhanced image.